Projects / Programmes
Translational oncology: development and validtion of immunogene therapy with Interleukin-12 combined with surgery for canine cancers of the oral cavity and skin
| Code |
Science |
Field |
Subfield |
| 3.04.00 |
Medical sciences |
Oncology |
|
| Code |
Science |
Field |
| B200 |
Biomedical sciences |
Cytology, oncology, cancerology |
| Code |
Science |
Field |
| 3.02 |
Medical and Health Sciences |
Clinical medicine |
translational oncology, gene therapy, dog's tumors, interleukin-12
Researchers (25)
Organisations (3)
Abstract
Scientific background and problem statement: In the last decades immunogene therapy for the treatment of cancer is gaining in importance. Currently there are 59 running immunegene studies in the field of human oncology, 11 of them are using plasmid DNA encoding the cytokine interleukin 12 (IL-12) indicating the great potential of this kind of therapy. In spite of marked advances in the field of gene therapy, the problem of safety and efficiency of this therapy in combination with standard treatment modalities remains. In the process of translation of these therapies to human medicine, the importance of clinical studies in veterinary medicine is becoming more and more valued, especially in dogs, namely spontaneously occurring tumors in dogs and the similarity of canine and human immune systems represent an important translational bridge accelerating the transition to human medicine.
The aim of the proposed project is the development and validation of a safer adjuvant immune gene therapy with IL-12 in combination with surgery for the treatment of oral and skin tumors in dogs. For this purpose plasmid DNA encoding canine IL-12 gene under transcriptional control of the tissue specific promoter for skin will be constructed. Plasmid will be evaluated in cell cultures and in tumor models in vivo. Clinical protocol will be prepared, and all necessary approvals will be obtained from the competent authorities and first patients will be recruited into the clinical study.
Methods: Plasmids encoding canine IL-12 under the transcriptional control of a tissue specific promoter and with kanamycin resistance gene will be prepared using standard molecular biology methods. Plasmid without the antibiotic resistance gene will be prepared according to the instructions of the supplier of ORT® technology. Appropriate bacterial strains will be transformed with the plasmids to produce larger quantities of endotoxin free plasmid to be used in further experiments. The expression profiles of the plasmids will be determined in different canine and human cells using RT-PCR and ELISA. Additionally proliferation assay will be performed. Antitumor effect of the prepared plasmids will be determined in canine tumor xenografts in nude mice. Antitumor effectiveness will be assessed by tumor growth delay assay and tumor control probability assay and histological analysis of the tumors will be conducted. Clinical study will be executed at the Clinic for Surgery and Small Animal of the Veterinary faculty University of Ljubljana.
Execution and management of the project: The work on the project will be executed at the laboratories of 5 partners participating in the project, where all necessary equipment for molecular biology, cell cultures, immunocytochemistry, animal experiments and clinical study is available. The researchers involved in the project have vast experience in the field of plasmid preparation, cell culture isolation, electroporation as a drug and gene delivery method, clinical protocols preparation, and electroporation in veterinary patients.
The management and coordination of the project will be assured by regular meetings and constant communication with partners on the project.
Relevance and potential impact of the results: The proposed project is designed as a preclinical and clinical project that addresses important issues regarding safe use of cancer gene therapy in clinical setting. The results of the project, if successful, may have impact on implementation of gene therapy in veterinary oncology and above all the translation of such clinical studies into human oncology in Slovenia and worldwide.
Significance for science
The development of recombinant technologies for the production of recombinant human cytokines hold a great promise for cancer treatment, as our immune system is very effective in combating cancer cells. Unfortunately, the clinical trials using recombinant proteins, such as interleukins and interferons, were not successfully completed, due to deleterious side effects. Immunogene therapies with targeted delivery of gene coding for cytokines resulted in the levels of cytokines that are more constant and lower compared to recombinant proteins. Therefore, according to preclinical studies and early results of some clinical studies, demonstrating good antitumor effect without undesired side effects, these therapies hold great promised for cancer treatment. In contrast to biological agents, that target single molecules or pathway in cancer cells, immune therapies stimulate organisms’ immune system to systemically combat cancer. Especially, combined with local standard cancer treatment, such as surgery and radiotherapy, these therapies could add safe a systemic component to the cancer treatment. Although, gene therapy approaches are numerous, nonviral vectors, i.e. plasmid DNA, are evaluated in more and more clinical trials. The use of electroporation for increased delivery of plasmid DNA already proved its applicability in clinical trials. Our proposed project aims at development of safe plasmid vectors, evaluation of the vectors combined with electroporation in preclinical study and testing its applicability for the treatment of spontaneous tumors in dogs in combination with surgery. Therefore, the results of our project will bring new basic knowledge in the construction and preparation of the vectors on one side and translate the therapy into veterinary oncology, facilitating the way also in human clinical trials.
Significance for the country
Despite the fact that there are currently a lot of research efforts, as well as money focused on cancer prevention, early diagnostics and biomarkers, translational research in the field of the development of new therapies is still topical. Within the project we developed a new safe immunogenic therapy, which we tested on spontaneous tumors of dogs. Our results have shown that combined therapy has led to the successful treatment of dog tumors. Therapy itself is more affordable than, for example, chemotherapy, and thus leads to reduced owner costs. In addition, the implementation of therapy in veterinary oncology led to the preparation of a study in human oncology, where the need for new successful therapies, especially in metastatic melanoma, is required. We also filed a patent application for the newly made plasmid. The project results were repeatedly published not only as scientific articles, but also as contributions to many international conferences. Last but not least, within the project, workshops were also organized by researchers and veterinarians from all over Europe, as well as other countries, such as Brazil. It is also a notible fact that included dogs in the clinical study were not only from Slovenia, but also from neighboring countries. All the above facts point to the excellent reputation of our research group and of Slovenia around the world.
Most important scientific results
Annual report
2014,
2015,
final report
Most important socioeconomically and culturally relevant results
Annual report
2014,
2015,
final report
